First Report of Predation by a Stink Bug on a Walking-Stick with Refections on Evolutionary Mechanisms for Camoufage

Jane Margaret Costa de Frontin Werneck (  [email protected] ) Instituto Oswaldo Cruz https://orcid.org/0000-0001-6664-1394 Lucas Torres Fundacao Oswaldo Cruz David Willian Provance Fundacao Oswaldo Cruz Ricardo Brugnera Universidade Federal do Rio Grande do Sul Jocelia Grazia Universidade Federal do Rio Grande do Sul

Research note

Keywords: , Asopinae, predation, camoufage

Posted Date: July 2nd, 2019

DOI: https://doi.org/10.21203/rs.2.10812/v1

License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License

Page 1/11 Abstract

Objective: The stink bug Supputius cincticeps is a well-known neotropical, generalist predator. However, in contrast to other predators, S. cincticeps also need to feed on plants to complete its life cycle. The aim of this report is to describe predation by S. cincticeps on Cladomorphus phyllinus, a walking-stick insect that feeds on leaves of several fruit trees, and is considered one of the largest walk-stick insect in the Southern Hemisphere. Also a new hypothesis for camoufage is discussed. Results: Suputius cincticeps, recently brought from nature, was accidentally introduced into a colony of C. phyllinus, while feeding them with guava leaves. Once the nymph of S. cincticeps detected the C. phyllinus female it displayed immediately feeding behavior. Both adults and nymphs of C. phyllinus were observed to be predated by S. cincticeps. This new predatory interaction offers a novel evolutionary hypothesis for camoufage based on horizontal transfer of genes a process that might be occurring in nature. Keywords: Phasmatodea, Asopinae, predation, camoufage

Introduction

The predatory stink bugs ( Asopinae) is the only group of Pentatomidae that needs to feed on other to complete their life cycle, contrasting with other groups of the that are mostly phytophagous [1, 2]. Asopinae is considered and recognized as a natural taxon, distributed worldwide and comprising approximately 63 genera and 303 species [3, 4]. Predatory stink bugs are also important tools in biological control: several species are used to control populations of other in agricultural systems, reducing need for some pesticides [2, 5, 6].

The predatory stink bug Supputius cincticeps (Stål, 1860) is a well-known, generalist predator that feeds mostly on larvae of Lepidoptera [7] and other insects under lab conditions [8]. However, in contrast to other predators, S. cincticeps also need to feed on plants to complete its life-cycle (R. Brugnera, unpublished data). This insect inhabits several ecosystems, and is widely distributed across the Neotropical region [9]. Supputius cincticeps is considered important in biological control in crops and forests, particularly in Brazil, which has led to detailed studies of its biology, behavior, morphology and reproduction [6, 2, 10].

Cladomorphus phyllinus Gray, 1835 is a walking-stick insect ( Phasmatodea). The Order includes 13 families, 523 genera and 2.822 species. Of these species, 591 occur in the Neotropical region [11], and more than 200 in Brazil [12]. Cladomorphus phyllinus feeds on leaves of several fruit trees, and is considered one of the largest walk-stick insect species in the Southern Hemisphere, where female can reach 22 cm long [13]. The entire group is little studied [14, 15], particularly the South American species [11].

Presenting a biological cycle of around 280 days from egg to adult, C. phyllinus can be reared under conditions of captivity [16].

Page 2/11 Materials And Methods

In 1917, we captured four females and two males, from secondary fragments of the Atlantic Forest near Petrópolis, Rio de Janeiro, Brazil (22º 30' 18" S; 43º 10' 43" W; 809m); these individuals were used to establish a robust colony of more than 200 individuals. In order to record bionomical aspects, individuals are being kept in captivity. The colonies are being maintained under environmental conditions ranging in temperature from 15-32°C and 65-93% relative humidity. Adult specimens are kept in enclosures housing from 10 to 20 individuals and provided with leaves from Psidium guajava (guava trees) as a primary food source.

The C. phyllinus specimens mentioned in this paper were identifed [17, 11] and deposited in the collection Jane Costa & Lima Neiva of the Oswaldo Cruz Institute Entomological Collection , Fiocruz [18, 19].

Results

While providing recently collected guava leaves to one of the colonies, a ffth instar nymph of S. cincticeps was introduced accidentally into the one of the enclosures housing 20 adult female of C. phyllinus. Once the nymph detected an adult female of C. phyllinus, it began immediately to display feeding behavior. It frst chose to attempt feeding at the pulvilo, which resulted in the repulsive reaction of the adult female showing some sensitivity, manifested as movements of the leg in a manner for scraping off as irritant (Fig 1A). The S. cincticeps nymph persisted, appeared to succeed at sucking the pulvilo for approximately 3 min. The S. cincticeps nymphmoved to another region of the leg and commenced to feed on the mid-tibia (Fig 1B).

The asopine nymph was then transferred to a small pot together with fve frst-instar nymphs of C. phyllinus, along with some leaves from Calliandra brevipes Benth,1840, a food source C. phyllinus nymphs. Two days later, S. cincticeps was recorded feeding on C. phyllinus nymphs; S. cincticeps nymph fed upon all fve nymphs of C. phyllinus over the succeeding nine days, leaving the completely dry, dead bodies of the C. phyllinus nymphs in the bottom of the pot. The nymph of S. cincticeps did not survive to adulthood: died three days after its last meal, feeding on C. phyllinus frst-instar nymphs (Fig 2)

Discussion

This report is the frst to document predation of S. cincticeps on C. phyllinus. Supputius cinctipes is known for its eclectic feeding behavior, attacking a variety of insects. It has been evaluated carefully as a potential candidate for natural control of agricultural pests. This group of insects can also feed on plant sap when insect prey is scarce [20-23]; this alteration in food source, referred to as zoophytophagous behavior, facilitates increased survival of nymphs and extends the longevity and fecundity of adult insects [24, 25]. Some species need plants for completing the developmental cycle. Supputius cincticeps following the rule, dies in the nymphal stage if some vegetal is not available for feeding.

Page 3/11 These stink bugs can be phytozoophagous or zoophytophagous: phytozoophagous insects are herbivores that occasionally feed on small insects; and vice versa for zoophytophagous insects [21, 26]. These feeding habits elucidate how pentatomid predators use different food sources as well as ecological and evolutionary changes in feeding [27, 28]. The zoophytophagous S. cincticeps can be an important component of biological control programs of soybean pests, and pests of other beans, and cotton in Brazil [29], especially pest species of Lepidoptera, Coleoptera, Diptera, and Hemiptera [30- 32]. They apparently see improved development and reproduction of zoophytophagous predators when they feed on both plants and insects [33].

Possible mechanisms into how morphological changes arise that provide camoufage to insects for escaping predators have long been discussed [34]. The most prevalent concept is that new morphologies evolved through the traditional combination of germ-line mutations and natural selection [35]. However, one can easily speculate that other mechanisms could be used to generate different camoufage strategies. Give the diverse predatory feeding behavior of S. cincticeps, we propose that the camoufage of C. phyllinus could be facilitated by horizontal or lateral transfer of plant-derived genetic material leading to development of a form resembling a tree stem.

While unprecedented and speculative, a hypothesis based on transfer of genetic material is plausible since the predatory Pentatomidae feed by inserting their stylet into the body of the prey, and injecting toxins, enzymes, or both, before sucking prey tissues [30, 36]. Asopines are known to use the plant juices for producing saliva, which will be used for predation activity [1]. This behavior could also provide a conduit by which to introduce plant genes, or any bacteria or virus that coexists with the plant.

Horizontal transfer of genes was frst reported in 1928 [37] and, is now accepted as an important mechanism for evolutionary processes. Horizontal transfer is known to have occurred among prokaryotes [ 38, 39] and the phenomenon appears to have had signifcance for unicellular eukaryotes as well [40]. Still, the prevalence and importance of horizontal transfer in the evolution of multicellular- eukaryotes remain unclear [41]. Some clear examples have been documented, such aspea aphids (Acyrthosiphon pisum) which have multiple genes from fungi [42]. Plants, fungi, and microorganisms can synthesize carotenoids, but torulene made by pea aphids is the only carotenoid known to be synthesized by an organism in the kingdom [43]. Also, HhMAN1, a gene in the genome of the coffee borer beetle (Hypothenemus hampei), resembles bacterial genes, and is thought to be transferred from bacteria in the beetle's gut [44].

Here we report a generalist predator S. cincticeps feeding on C. phyllinus, the latter of which presents a camoufage in the form of an intriguing phenotype, the “walking stick” body form. We propose a novel hypothesis: that the camoufage could have been arisen via of a plant morph gene to the insect genome by an insect vector.

Deep molecular studies at the level of genome sequencing are needed to test this hypothesis opening new models and interesting possibilities in the universe of the genome interactions and dynamics.

Page 4/11 Limitations

-The new interaction between a stink bug and a walk-stick insect here reported was not recorded in natural environment. However, S. cincticeps presented its predatory behavior immediately after its accidental introduction in one of the enclosures, housing C. phyllinus specimens.

- The report of the predation by S. cincticeps on C. phyllinus offer a new evolutionary hypothesis on camoufage based on horizontal transfer of genes however, genome sequencing analyses to test this hypothesis are required.

Declarations Acknowledgments

We are grateful to Instituto Chico Mendes de Conservação da Biodiversidade for issuing collecting permits. We would like to thank Dr. Andrew Townsend Peterson for reviewing the text and for valuable comments and suggestions; Gustavo de Frontin Werneck for helping the maintenance of the insect colonies; CNPq for fnancial support.

Authors’ contributions

JC identifed walking-stick insect, kept the insect colonies, recorded the predation and wrote the paper. LC reared the insects, wrote the paper. DWPJr wrote the paper. RB and JG identifed stink bug and wrote the paper. All authors read and approved the fnal manuscript.

Funding

JC is recipient for a Research Productivity Fellowship from Brazilian National Council for Scientifc and Technological Development.

Availability of data and materials

Studied specimens were deposited at the collection Jane Costa & Lima Neiva of the entomological collection of the Oswaldo Cruz Institute, Fiocruz, Brazil.

Ethics approval and consent to participate

Not applicable.

Page 5/11 Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Author details

1Laboratory of Entomological , Oswaldo Cruz Institute, Fiocruz, Av. Brasil, 4365 Manguinhos, Rio de Janeiro, Brasil, Cep 21040-360. 2Center of Technological Development in Health (CDTS)/National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN), FIOCRUZ, Rio of Janeiro, RJ, Brazil . 3Laboratory of Entomology Systematics, Department of Zoology & Graduate Program on Animal Biology, Federal University of Rio Grande do Sul, Brazil.

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Figures

Page 9/11 Figure 1

Predation by a ffth-instar nymph of Supputius cincticeps (Stål, 1860) on a female of Cladomorphus phyllinus Gray, 1835: A- feeding the pulvilo; B- feeding the mid-tibia.

Page 10/11 Figure 2

Predation by a ffth-instar nymph of Supputius cincticeps (Stål, 1860) on a frst instar nymph of Cladomorphus phyllinus Gray, 1835:

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